The Inner Nuclear Membrane Protein Nemp1 Is a New Type of RanGTP-Binding Protein in Eukaryotes
Fig 2
Oligomerization of Nemp1 through the TMs.
A. Co-IP of Xl_Nemp1 with Xl_Nemp1 itself, MAN1, or Emerin. Xl_Nemp1-HA mRNA was coinjected into the animal pole region of two cell stage Xenopus embryos with mRNA for Xl_Nemp1-Myc, XMAN1-Myc, or Hs_emerin-Myc. Injected embryos were collected at the late blastula stage (stage 9) and lysed with lysis buffer A. Black arrowheads, expected product bands; white arrowheads, immunoglobulin bands; asterisks, shifted bands of Emerin due to phosphorylation [40]. B. Co-IP of Nemp1 with its deletion constructs. mRNA for Xl_Nemp1-HA was injected into Xenopus embryos with mRNA for deletion constructs of Xl_Nemp1-Myc. Deletion constructs of Nemp1, ΔN, ΔA, ΔTM, ΔB, N, SP+A, SP+B, and Ct (Δ, deleted; N, the N-terminal region; A, region A; TM, transmembrane domains; B, region B; SP, signal peptide; Ct, the C-terminal region; S2 Fig for diagrams) were used (see [10] for more detail). After immunoprecipitation against Myc, western blotting was performed with anti-Myc or-HA antibody as indicated below each panel.